Advertisement

Springer Nature is making Coronavirus research free. View research | View latest news | Sign up for updates

The role ofSaccharomyces cerevisiae Cdc40p in DNA replication and mitotic spindle formation and/or maintenance

  • 66 Accesses

  • 40 Citations

Abstract

Successful progression through the cell cycle requires the coupling of mitotic spindle formation to DNA replication. In this report we present evidence suggesting that, inSaccharomyces cerevisiae, theCDC40 gene product is required to regulate both DNA replication and mitotic spindle formation. The deduced amino acid sequence ofCDC40 (455 amino acids) contains four copies of a β-transducin-like repeat. Cdc40p is essential only at elevated temperatures, as a complete deletion or a truncated protein (deletion of the C-terminal 217 amino acids in thecdc40-1 allele) results in normal vegetative growth at 23°C, and cell cycle arrest at 36°C. In the mitotic cell cycle Cdc40p is apparently required for at least two steps: (1) for entry into S phase (neither DNA synthesis, nor mitotic spindle formation occurs at 36°C and (2) for completion of S-phase (cdc40::LEU2 cells cannot complete the cell cycle when returned to the permissive temperature in the presence of hydroxyurea). The role of Cdc40p as a regulatory protein linking DNA synthesis, spindle assembly/maintenance, and maturation promoting factor (MPF) activity is discussed.

This is a preview of subscription content, log in to check access.

References

  1. Boeke JD, Lacroute F, Fink GR (1984) A positive selection for mutants lacking ortidine-5-phosphate decarboxylase activity in yeast. Mol Gen Genet 197:345–346

  2. Broek D, Bartlett R, Crawford K, Nurse P (1991) Involvement of p34cdc2 in establishing the dependency of S phase on mitosis. Nature 349:388–393

  3. Bueno A, Russell P (1992) Dual functions ofCDC6: a yeastprotein required for DNA replication also inhibits nuclear division. EMBO J. 11:2167–2176

  4. Byers B (1981) Cytology of the yeast life cycle. In: Strathern JN, Johns EW, Broach JR (eds) The molecular biology of the yeast Saccharomyces: Life cycle and inheritance. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, pp 59–97

  5. Chien CT, Bartel PL, Sternglanz R, Field S (1991) The two-hybrid system: a method to identify and clone genes for proteins that interact with a protein of interest. Proc Natl Acad Sci USA 88:9578–9582

  6. Dasso M (1993)RCC1 in the cell cycle: the regulator of chromosome condensation takes on new roles. Trends Biochem Sci 18:96–101

  7. Deng XW, Matusi M, Wei M, Wagner D, Chu AM, Feldman KA, Quail PH (1992)COP1, anArabidopsis regulatory gene, encodes a protein with both zinc-binding motif and a Gβ homologous domain. Cell 71:791–801

  8. Duronio RJ, Gordon JI, Boguski MS (1992) Comparative analysis of the β transducin family with identification of several new members includingPWP1, a nonessential gene ofSaccharomyces cerevisiae that is divergently transcribed fromNMT1. Proteins 13:41–56

  9. Dynlacht BD, Weinzierl ROJ, Admon A, Tjian R (1993) The dTAFII80 subunit of TFIID contains β-transducin repeats. Nature 363:176–179

  10. Enoch T, Carr AM, Nurse P (1991) Fission yeast genes involved in coupling mitosis to completion of DNA replication. Genes Dev 6:2035–2046

  11. Enoch T, Carr AM, Nurse P (1992) Fission yeast genes involved in coupling mitosis to completion of DNA replication. Genes Dev 6:2035–2046

  12. Enoch T, Nurse P (1991) Coupling M phase and S phase: controls maintaining the dependence of mitosis on chromosome replication. Cell 65:921–923

  13. Fitch I, Dahmann C, Surana U, Amon A, Nasmyth K, Goetsch L, Byers B, Futcher B (1992) Characterization of four B-type cyclin genes of the budding yeastSaccharomyces cerevisiae. Mol Biol Cell 3:805–818

  14. Foiani M, Marini F, Gamba D, Lucchini J, Plevani P (1994) The β subunit of the DNA polymerase α-primase complex inSaccharomyces cerevisiae executes an essential function at the initial stage of DNA replication. Mol Cell Biol 14:923–933

  15. Fong HKW, Hurley JB, Hopkins RS, Miake-Lye R, Johnson MS, Doolittle RF, Simon MI (1986) Repetitive segmental structure in the transducin β subunit: homology with theCDC4 gene and identification of related mRNAs. Proc Natl Acad Sci USA 83:2162–2166

  16. Gietz RD, Sugino A (1988) New Yeast-Escherichia coli shuttle vectors constructed within vitro mutagenized six-base pair restriction sites. Gene 74:527–534

  17. Giroux CN, Mis JR, Pierce MK, Kohalmi SE, Kunz BA (1988) DNA sequence analysis of spontaneous mutations in theSUP4-o gene ofSaccharomyces cerevisiae. Mol Cell Biol 8:978–981

  18. Goebl M, Yanagida M (1991) The TPR snap helix: a novel protein repeat motif from mitosis to transcription. Trends Biochem Sci 16:173–177

  19. Hartwell LH (1976) Sequential function of gene products relative to DNA synthesis in the yeast cell cycle. J Mol Biol 104:803–817

  20. Hartwell LH, Weinert TA (1989) Checkpoints: Controls that ensure the order of cell cycle events. Science 246:629–634

  21. Hinnebusch AG (1988) Novel mechanism of translational control inSaccharomyces cerevisiae. Trends Genet 4:169–174

  22. Hogan E, Koshland D (1992) Addition of extra origins of replication to a minichromosome suppresses its mitotic loss incdc6 andcdc14 mutants ofSaccharomyces cerevisiae. Proc Natl Acad Sci USA 89:3098–3102

  23. Hoyt MA, Totis L, Roberts BT (1991)S. cerevisiae genes required for cell cycle arrest in response to loss of microtubule function. Cell 66:507–517

  24. Hunter T, Pines J (1991) Cyclins and cancer. Cell 66:1071–1074

  25. Ito H, Fukuda Y, Murata K, Kimura A (1983) Transformation of intact yeast cells treated with alkali cations. J Bacteriol 153:163–168

  26. Jacobs CW, Adams AEM, Szaniszlo PJ, Pringle JR (1988) Functions of microtubules in theSaccharomyces cerevisiae cell cycle. J Cell Biol 107:1409–1426

  27. Jimenez G, Yucel J, Rowley R, Subramani S (1992) Therad3 + gene ofSchizosaccharomyces pombe is involved in multiple checkpoint functions and in DNA repair. Proc Natl Acad Sci 89:4952–4956

  28. Johnston GC, Singer RA (1980) Ribosomal precursor RNA metabolism and cell division in the yeastSaccharomyces cerevisiae. Mol Gen Genet 178:357–360

  29. Kassir Y, Kupiec M, Shalom H, Simchen G (1985) Cloning and mapping ofCDC40 aSaccharomyces cerevisiae gene with a role in DNA repair. Current Genet 9:253–257

  30. Kassir Y, Simchen G (1978) Meiotic recombination and DNA synthesis in a new cell cycle mutant ofSaccharomyces cerevisiae. Genetics 90:49–68

  31. Kassir Y, Simchen G (1991) Monitoring meiosis and sporulation inSaccharomyces cerevisiae. Methods Enzymol 194:94–109

  32. Koch C, Moll T, Neuberg M, Ahorn H, Nasmyth K (1993) A role for the transcription factors Mbp1 and Swi4 in progression from G1 to S phase. Science 261:1551–1557

  33. Lew DJ, Reed SI (1993) Morphogenesis in the yeast cell cycle: regulation by Cdc28 and Cyclins. J Cell Biol 120:1305–1320

  34. Li R, Murray AW (1991) Feedback control of mitosis in budding yeast. Cell 66:519–531

  35. Lisziewicz J, Godany A, Agoston D, Kuntzel H (1988) Cloning and characterization of theSaccharomyces cerevisiae CDC6 gene. Nucleic Acids Res 16:11507–11520

  36. Maniatis T, Fritsch EF, Sambrook J (1982) Molecular Cloning: a Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

  37. Matsumoto T, Beach D (1991) Thespil GTPase interacts withRCC1 in cell cycle dependency. Cold Spring Harbor Symp Quant Biol LVI:385–398

  38. Millar JBA, Russell P (1992) Thecdc25 M-phase inducer: an unconventional protein phosphatase. Cell 68:407–410

  39. Mukai Y, Harashima S, Oshima Y (1991)AAR1/TUP1 protein with a structure similar to that of the β subunit of G proteins, is required fora1-α2 and α2 repression in cell type control ofSaccharomyces cerevisiae. Mol Cell Biol 11:3773–3779

  40. Nasmyth K (1993) Control of the yeast cell cycle by the Cdc28 protein kinase. Curr Opin Cell Biol 5:166–179

  41. Nurse P (1990) Universal control mechanism regulating onset of M-phase. Nature 344:503–508

  42. Ogas J, Andrews BJ, Herskowitz I (1991) Transcriptional activation ofCLN1, CLN2, a putative new G1 cyclin (HCS26) bySWI4, a positive regulator of G1-specific transcription. Cell 66:1015–1026

  43. Pringle JR, Hartwell LH (1981) TheSaccharomyces cerevisiae cell cycle. In: Strathern JN, Jones EW, Broach JR (eds). The Molecular Biology of the YeastSaccharomyces. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, pp 94–142

  44. Reed SI (1991) G1-specific cyclins: in search of an S-phase promoting factor. Trends Genet 7:95–99

  45. Reed SI, Wittenberg C (1990) A mitotic role for the Cdc28 protein kinase ofS. cerevisiae. Proc Natl Acad Sci USA 87:5697–5701

  46. Richardson H, Lew DJ, Henze M, Sugimoto K, Reed SI (1992) Cyclin-B homologs inSaccharomyces cerevisiae function in S phase and in G2. Genes Dev 6:2021–2034

  47. Roitgrund C, Steinlauf R, Kupiec M (1993) Donation: a new, facile method of gene replacement in yeast. Mol Gen Genet 237:306–310

  48. Rose MD, Fink GR (1987)KAR1, a gene required for function of both intranuclear and extranuclear microtubules in yeast. Cell 48:1047–1060

  49. Rose MD, Misra L, Vogel JP (1989)KAR2, a karyogamy gene, is the yeast homolog of the mammalian Bip/GRP78 gene. Cell 57:1211–1221

  50. Rose M, Winston F, Hieter P (1990) Methods in Yeast Genetics. A Laboratory Course Manual. Cold Spring Harbor Laboratory Press. Cold Spring Harbor, New York

  51. Rothstein R (1991) Targeting, disruption, replacement and allele rescue: integrative transformation in yeast. Methods Enzymol 194:281–301

  52. Rowley R, Subramani S, Young PG (1992) Checkpoint controls inSchizosaccharomyces pombe: rad1. EMBO J 11:1335–1342

  53. Russel P, Nurse P (1986)cdc25 + functions as an inducer in the mitotic control of fission yeast. Cell 45:145–153

  54. Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain termination inhibitors. Proc Natl Acad Sci USA 74:5463–5467

  55. Schild D, Anathaswamy HN, Mortimer R (1981) An endomitotic effect of a cell cycle mutation inSaccharomyces cerevisiae. Genetics 97:551–562

  56. Schwob E, Nasmyth K (1993)CLB5 andCLB6, a new pair of B cyclins involved in DNA replication inSaccharomyces cerevisiae. Genes Dev 7:1160–1175

  57. Seino H, Nishitani H, Seki T, Hisamoto N, Tazunoki T, Shiraki N, Ohtsubo M, Yamashita K, Sekiguchi T, Nishimoto T (1991)RCC1 is a nuclear protein required for coupling activation ofcdc2 kinase with DNA synthesis and for start of the cell cycle. Cold Spring Harbor Symp Quant Biol LVI:367–376

  58. Sethi N, Monteagudo C, Koshland D, Hogan E, Burke DJ (1991) TheCDC20 gene product ofSaccharomyces cerevisiae, a β-transducin homolog, is required for a subset of microtubule-dependent cellular processes. Mol Cell Biol 11:5592–5602

  59. Sherman F (1991) Getting started with yeast. Meth Enzymology 194:3–21

  60. Surana U, Robitsch H, Price C, Schuster T, Fitch I, Futcher AB, Nasmyth K (1991) The role ofCDC28 and cyclins during mitosis in the budding yeastS. cerevisiae. Cell 65:145–161

  61. Tzamarias D, Struhl K (1994) Functional dissection of the yeast Cyc8-Tup1 transcriptional co-repressor complex. Nature 369:758–761

  62. Vallen EA, Scherson TY, Roberts T, Zee K, Rose MD (1992) Asymmetric mitotic segregation of the yeast spindle pole body. Cell 69:505–515

  63. Walworth N, Davey S, Beach D (1993) Fission yeastchk1 protein kinase links therad checkpoint pathway tocdc2. Nature 363:368–371

  64. Weinert TA, Hartwell LH (1988) TheRAD9 gene controls the cell cycle response to DNA damage inSaccharomyces cerevisiae. Science 241:317–322

  65. Weinert TA, Hartwell LH (1993) Cell cycle arrest ofcdc mutants and specificity of theRAD9 checkpoint. Genetics 134:63–80

  66. Weinert TA, Kiser GL, Hartwell LH (1994) Mitotic checkpoint genes in budding yeast and the dependence of mitosis on DNA replication and repair. Genes & Develop. 8:652–665

  67. Whiteway M, Hougan L, Dignard D, Thomas DY, Bell L, Saari GC, Grant FJ, O'Hara P, MacKay VL (1989) TheSTE4 andSTE18 genes of yeast encode potential β and τ subunits of the mating factor receptor-coupled G protein. Cell 56:467–477

  68. Williams FE, Trumbly RJ (1990) Characterization ofTUP1, a mediator of glucose repression inSaccharomyces cerevisiae. Mol Cell Biol 10:6500–6511

  69. Williams FE, Varanasi U, Trumbly RJ (1991) TheCYC8 andTUP1 proteins involved in glucose repression inSaccharomyces cerevisiae are associated in a protein complex. Mol Cell Biol 11:3307–3316

  70. Zhou C, Huang SH, Jong AY (1989) Molecular cloning ofSaccharomyces cerevisiae CDC6 gene. J Biol Chem 264:9022–9029

Download references

Author information

Additional information

Communicated by C. Hollenberg

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Vaisman, N., Tsouladze, A., Robzyk, K. et al. The role ofSaccharomyces cerevisiae Cdc40p in DNA replication and mitotic spindle formation and/or maintenance. Molec. Gen. Genet. 247, 123–136 (1995). https://doi.org/10.1007/BF00705642

Download citation

Key words

  • CDC40
  • DNA replication
  • Mitotic spindle assembly
  • cyclins
  • Saccharomyces cerevisiae